What is the application and preparation method of HEPES buffer? CAS NO.7365-45-9

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 Hydroxyethylpiperazine ethanesulfonic acid HEPES is a non-ionic amphoteric buffer whose main component is hydroxyethylpiperazine ethanethiosulfonic acid. It has good buffering capacity in the range of PH7.2-7.4. Hepes is widely used as a biological buffer as a buffer reagent in cell culture media, as a buffer and eluent in protein research, as well as in DNA/RNA extraction kits and PCR diagnostic kits.

One of the great things about Hepes as a buffer is that it can maintain a relatively constant pH value in open culture or cell observation. Under such culture conditions, the cell culture vessel needs to be closed and perfect to avoid the small amount required in the culture medium. Carbonates are dispersed into the air. There are two ways to use Hepes buffer. details as follows:
1. Hepes can be directly added to the prepared culture solution according to the required concentration, and then sterilized by filtration. Add 2.38 grams of HEPES to every 1000ml of culture solution, adjust the pH to 7.2 with 1N NaOH after dissolving, filter and sterilize before use .At this time, the concentration of HEPES used is 10 mmol/L.
2. It can also be made into 100 x stock solution (1 mol/L). Before use, add 99 ml of culture medium to 1 ml of stock solution, and the final application concentration is still 10 mmol/L. l mol/L (100 x) HEPES stock solution preparation method: Dissolve 23.8 g of HEPES in 90 ml of double-distilled water, adjust the pH to 7.5-8.0 with 1N NaOH, then dilute to 100 ml with water, filter and sterilize, and divide into vials ( 2ml/bottle), need to be stored at 4°C or -20°C.
In daily biochemical experiments, especially in the process of cell culture, Hepes buffer is usually indispensable. The use of Hepes buffer has been understood, and then we will learn about the preparation method of Hepes.
The preparation method of Hepes: directly use 1,2-dichloroethane as solvent, add hydroxyethylpiperazine (5.00g, 0.02mol), potassium carbonate K2CO3 (6.00 g, 0.04 mol), 50 mL of 1,2-dichloroethane, heated in an oil bath at 90 °C (boiling point of 1,2-dichloroethane 85 °C), and stirred for 20 h. The reaction was stopped, filtered, and the filtered salt was washed with 200 mL of ethyl acetate (EA). The filtrate was spin-dried to obtain 2.6 g of HEPES solid.
In summary, Hepes buffers are widely used, and the methods of use and preparation are not complicated. As a result, there are many Hepes merchants in the market, and there are actually only a few real domestic manufacturers. Therefore, the market is currently a mixed bag, and there are many middlemen who earn the difference. We recommend a manufacturer here, Hubei Xindesheng Materials Co., Ltd. Desheng has been specialized in the production and development of biological buffers for many years. It has multi-storey workshops and imported equipment, and has created a professional team responsible for R&D and production management, with high production efficiency. , The product quality is excellent, the warehouse stock is sufficient, and the order is shipped. If you are interested in learning about Desheng's products, you can contact the customer service on the official website of Desheng for details.